Mouse D-Dopachrome Tautomerase ELISA Kit

From Antibodies.com, this sandwich enzyme-linked immunosorbent assay (sELISA) is designed for the in vitro quantitative determination of mouse D-Dopachrome Tautomerase (DDT) in tissue homogenates, cell lysates and other biological fluids. The assay is run on a pre-coated 96-well microplate (12 x 8 well strips) and read colorimetrically at 450 nm on a standard microplate reader. Sensitivity is <0.141 ng/ml and the measuring range is 0.312–20 ng/ml, with a total assay time of 4 hours 30 minutes.

In the sandwich format, an antibody specific for DDT is pre-coated onto the plate. Standards and samples are added and any DDT present is captured by the immobilized antibody. After washing, a biotinylated anti-DDT antibody binds the captured analyte, followed by HRP-Avidin conjugate. TMB substrate produces a blue colour that turns yellow after addition of the acidic stop solution; the intensity of yellow is proportional to DDT concentration and is read against a standard curve.

Average recovery in serum (n=5) is 91% (range 80–102%), in EDTA plasma 90% (range 81–100%) and in heparin plasma 84% (range 80–89%). Dilutional linearity across 1:2 to 1:16 dilutions is reported for serum (87–91%, 87–107%, 74–101%, 92–97%), EDTA plasma (90–105%, 84–101%, 90–101%, 79–108%) and heparin plasma (84–95%, 92–105%, 82–105%, 89–91%), n=5 each. The kit includes a pre-coated 96-well microplate, lyophilized standard, detection solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers, with components stored at +4°C or -20°C as instructed.